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FIGURE 4 NB-5-MT (2) suppresses hypoxia-mediated angiogenesis in vitro. (A-B) Under hypoxia, HCT116 cells were incubated with 1 mM of melatonin (M) or 2 for 16 hr Conditioned medium (CM) was collected from HCT116 cells or MDA-MB-231 cells and added to HUVECs for angiogenesis assays. (A) Inhibitory effect of 2 on angiogenesis in human umbilical vein endothelial cells (HUVECs) in vitro. Representative images of tube formation (top), invasion (middle), and migration (bottom) assays from 3 independent experiments. (B) Relative anti-angiogenic effect of 2 using the conditioned media of HCT116 cells. Graphs of in vitro tube formation (left), invasion (middle), and migration (right). (C) Relative anti-angiogenic effect of 2 using the conditioned media of MDA-MB-231 cells. Graphs of in vitro tube formation (left), invasion (middle), and migration (right). Data are the mean ± SD from 3 independent experiments. *P <.05 and **P <.01, vehicle vs. hypoxia; +P <.05 and ++P <.01, hypoxia vs. hypoxia plus compound treatment